Secretory Expression and Characterization of Chinese Narcissus GNA-Like Lectin in Pichia pastoris
Narcissus tazetta lectin (NTL) is a GNA-like lectin, which has a wide potential application in medicine, agriculture, and glycobiology. In the present paper, the codon-optimized ntl gene was transformed into the yeast Pichia pastoris ; SDS–PAGE gel and western blotting analysis revealed that the rec...
Gespeichert in:
Veröffentlicht in: | Applied biochemistry and biotechnology 2017-08, Vol.182 (4), p.1433-1443 |
---|---|
Hauptverfasser: | , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
Zusammenfassung: | Narcissus tazetta
lectin (NTL) is a GNA-like lectin, which has a wide potential application in medicine, agriculture, and glycobiology. In the present paper, the codon-optimized
ntl
gene was transformed into the yeast
Pichia pastoris
; SDS–PAGE gel and western blotting analysis revealed that the recombinant lectin was expressed successfully in
Pichia
yeast. The similarity between the recombinant NTL and the native NTL was confirmed by circular dichroism (CD) and hemagglutination assay further. In the 5-L scale fermentator, the protein yield was as high as 1.2 g/L after fermentation for 96 h. In addition, the effect of metal ions (K
+
, Mg
2+
, Ca
2+
, and Cu
2+
), acid, and alkaline on hemagglutinating activity of NTL was tested, which provided biochemical characterizations of the mannose-binding lectin from Chinese
Narcissus
. |
---|---|
ISSN: | 0273-2289 1559-0291 |
DOI: | 10.1007/s12010-017-2408-5 |